β-hydroxybutyrate improves convulsions in mice carrying the GRIN1 Y647S/+ pathogenic variant

GRIN1 -related neurodevelopmental disorder ( GRIN1 -NDD) is characterized by clinically significant variation in the GRIN1 gene, which encodes the obligatory GluN1 subunit of N-methyl-D-aspartate receptors (NMDARs). The identified p.Tyr647Ser (Y647S) variant is carried by a 34-year-old female with seizures and intellectual disability. This study builds upon initial in vitro investigations of the functional impacts of this variant in the SYTANLAAF domain of the GluN1 M3 helix and examines its in vivo consequences in a mouse model.

To investigate in vitro functional impacts of NMDARs containing GluN1-Y647S variant subunits, GluN1-Y647S was co-expressed with wildtype GluN2A or GluN2B subunits in Xenopus laevis oocytes and HEK cells. Grin1 ^Y647S/+ mice were created by CRISPR-Cas9 endonuclease-mediated transgenesis and the molecular, electrophysiological, and behavioural consequences of the variant were examined. Additionally, de-identified patient data were collected to examine the representative nature of Grin1 ^Y647S/+ mice in modelling specific aspects of patient symptomology.

In vitro , NMDARs containing GluN1-Y647S showed altered sensitivity to endogenous agonists and negative allosteric modulators, and reduced cell surface trafficking. Ex vivo , Grin1 ^Y647S/+ mice displayed a reduction in whole brain GluN1 levels and a deficiency in NMDAR-mediated synaptic transmission in the hippocampus. Behaviourally, Grin1 ^Y647S/+ mice exhibited altered vocalizations, muscle strength, sociability, and problem-solving, as well as spontaneous convulsions that were ameliorated with supplementation of β-hydroxybutyrate (BHB), an endogenously produced ketone body.

The Y647S variant confers a complex in vivo phenotype, which reflects largely diminished properties of NMDAR function. As a result, Grin1 ^Y647S/+ mice display atypical behaviour in domains relevant to the clinical characteristics of GRIN1 -NDD and the individual carrying the variant, which allowed for the identification of BHB supplementation as a potential anti-convulsant treatment. Ultimately, the characterization of Grin1 ^Y647S/+ mice accomplished in the present work, expands our understanding of the mechanisms underlying GRIN1 -NDD and provides a foundation for the continued development of novel therapeutics.